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Description
Mouse monoamine oxidase,MAO ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect specimens using EDTA or heparin as an anticoagulant and centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 3. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-Test Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP enzyme conjugate are sequentially added to microwells pre-coated with monoamine oxidase and MAO capture antibodies. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the monoamine oxidase and MAO content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse monoamine oxidase,MAO ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Monoamine oxidases (MAO) are a family of enzymes that catalyze the oxidation of monoamines, using oxygen to remove their amine groups. They are found bound to the outer mitochondrial membrane of most cell types in the body. The first such enzyme was discovered in the liver by Mary Bernheim in 1928 and named tyramine oxidase. MAO belongs to a family of proteins called flavinamine oxidoreductases. They are important in breaking down monoamines ingested through food and also play a role in inactivating monoamine neurotransmitters. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, cell culture supernatant |
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4.8 ★★★★★
Based on 28 reviews
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Product Reviews
★★★★★ 5
Must must must
Must get! My dogs are obsessed with these! We go through them regularly!!
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Reviewed in the United States on January 6, 2026
★★★★★ 5
If Your Dog Can Rip The Hood Off Your Car--This Is The Dental Toy For Him/Her
I am always curious about buying the "number one best seller" that Amazon lists in any product category. And when it's an add-on item offered at a reasonable cost, I'm even more curious.
From reading the reviews, it appears that some dog owners ordered this item and were disappointed in it for a number of reasons, primarily because the 'hardness' of the toy caused dental problems. Obviously no one wants to buy a dental toy that actually causes teeth problems. However, some small breed owners must not be aware that their particular breed is subject to dental issues regardless of what toys they chew. Just off the top of my head, this type of problem often occurs with Yorkies, Chihauhaus, and Chinese Cresteds. You can use virtually any dental care method in existence and still watch your Chinese Crested's teeth fall out.
So small breeds shouldn't be given tough Nylabone toys like this Dinosaur model. If you watch any dog chew on this particular toy, you will quickly notice how small his/her teeth are in relation to the toy, and you will probably be surprised by the leverage and power he/she puts into each bite. They gnaw, they gnash, they continue to work at it until those tiny teeth lose the battle against the tough Nylabone. Comparing this to human teeth, everyone knows someone who fractures/cracks a tooth somehow. That's because our front "chicklet" teeth bite down on something hard, get broken from falling on your face, getting hit in the mouth, having teeth weakened by cavities, and from not seeing a dentist/hygienist on a regular basis--usually because we're afraid of a little pain.
We're always looking for relatively tough dental toys for our 182 pound Cane Corso named Dante. He tears apart any toy we buy him, usually in a matter of minutes. But this particular Nylabone Dura Chew has survived for months now. If you look at the size of his teeth, length and thickness, you will see a dog with the teeth and jaw muscles strong enough to tear the hood off your car--which is exactly what he did to our Prius. So if your dog is capable of ripping hoods off cars, this treat should be good for your pet monster. Just keep an eye on the toy so that you can toss it before it breaks down and is swallowed.
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Reviewed in the United States on March 10, 2015
★★★★★ 5
Best Nylabone Fido & I Agree
My dogs love these Dino Nylabone's. Over the years I have bought at least 30 of these nylabones for 3 different dogs. Two of which have loved the Dino shaped Nylabones the best. The third loves the nylabone daily dental bone the best but the dino is a close second. The T-Rex is their favorite but they like all dino shapes. Depending on the dog's chewing mood these have lasted anywhere from a month to 6 months before needing replacing. My super chewing husky will chew one up pretty quickly in about a month, my moderate chewing pit bull would take 3-4 months and my slow light chewing shiba inu would take 6 or more months. I typically have 2-3 chewable items per dog (it helps prevent toy hoarding and aggression) and these dino toys are always among the favorites.
*These are chewing style nylabones they are not supposed to be consumed quickly as a snack or treat. I consider these like a hobby or activity for my dogs to do not something they eat or a toy they play with. Chewing is a natural part of dog behavior and is good for dental hygiene. Giving your dog items they are supposed to chew on is a good way to prevent your dog from chewing on things he isn't supposed to.*
These are the white nylabone material which is a tougher material than the beige color nylabones. The white nylabones should be used for aggressive chewers. Beige nylabones are good for light chewers.
The T-Rex is about 6" by 5" the Long neck is about 3.5" by 7 inches and the Stegosaurus is about 6.5" by 4" they all seem to last about the same amount of time.
I can usually get them for $4-5 on amazon which makes them one of the best deals for a chewing nylabone which is another huge plus.
Overall My dogs and I love the Dino Nylabones and I will buy them as long as they are made. Great Product
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Reviewed in the United States on December 2, 2015
★★★★★ 4
Dog loves it and it's very durable, but not indestructible
I have a dog with very strong jaws. She's only thirty-five pounds but she goes through most toys like they're bubblegum. Even as a small puppy, she would destroy anything and everything. By the age of five or six months we had to give up on soft and rope toys altogether, and at a year old, she is limited to only hard chews like Kong, Nylabone, etc. Even then, she destroys most of these with relative ease. She's gone through chews others have recommended as long-lasting in literally ten minutes. The only toy she has that has lasted more than a month or so is her original red Kong, which she mostly licks instead of chews.
I initially bought the dinosaur for my other, less toy-murderous dog, but the power chewer quickly stole it from him. She was in love. She carried it everywhere and chewed on it constantly. I'm not sure if it's the shape or the texture she likes, or some combination thereof, but whatever the case, it instantly became her favorite. Even the most durable hard toys normally only last a few weeks with her, but her dino is going on two or three months now. It's chewed beyond recognition, all the nubs worn down, and it's very rapidly nearing the end of its life because little bits of plastic are falling off, but it still lasted far longer than most toys do. A few weeks ago I bought her a second one, in a different shape, and she's equally as enamored with it. All in all, it's not going to be indestructible if you have a power chewer, but it certainly lasts a heck of a lot longer than most!
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Reviewed in the United States on July 23, 2012
★★★★★ 5
Tough enough to Survive the big, bad CHARLIE!
I purchased this for my 80 pound yellow lab, Charlie.
Charlie is a very powerful chewer. Toys typically last about 2.3 seconds in his grasp.
Since Charlie's toy budget had ballooned to a point where I had to choose whether to fulfill his appetite for destruction, or pay the mortgage, I knew something had to change.
This is how I came across this nylabone chew. It appeared tough enough that charlie may get at least an hour of play from this toy before it was destroyed. The price was reasonable enough that it fit within what was left of the depleted toy budget. So I bought it.
Two quick days later, and the box arrived. I gave the package to Charlie, and told him it was another new toy for him to destroy. The smell of the nylabone vented through the cardboard, and he started to wag. He tore open the box and found the bone inside. At this point, he is elated. I removed the packaging and told him to go at it.
That was weeks ago, and the dinosaur bone still has yet to be defeated. Sure, it is missing a few spines, and his tail is heavily chewed, but the bone is not yet destroyed. That isn't from a lack on trying on Charlie's part. He chews on the bone almost daily, and hasn't gotten tired of it yet!
If you have a big dog who's toy budget is getting out of control, buy this bone! Small bits will come off here and there, so if your dog likes to eat what he destroys, keep an eye on him. However, all of the small bits that have come off Charlie's bone thus far are small enough that they should pass no problem.
A great bone, and if they release similar bones in the future, Charlie will be a happy customer.
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Reviewed in the United States on October 24, 2014